Seasonal serum concentrations of melatonin in cycling and noncycling mares.
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To determine whether secretory patterns of melatonin change throughout the seasons in mares, blood samples were drawn byvenipuncture from nine mares at noon and midnight for five successive days at monthly intervals from August through July at the University of Missouri in Columbia, MO. In addition, during September, December, March, and June, blood samples were drawn from indwelling catheters at 2-h intervals for 48 or 72 h. Mares were predominantly Quarter Horses weighing approximately 450 kg and ranged from 3 to 12 yr of age. Mares were housed in outdoor paddocks with three-sided run-in sheds for shelter. During the noon and midnight bleeding period, mares were placed in a larger open-sided barn with outside runs. Mares remained outdoors with the barn being used as a shelter in the event of inclement weather. All lights in the shed were converted to red light. Often, moonlight provided enough illumination to collect blood samples. Mares were returned to their normal paddock after each sampling period. For analysis of data, a mare was considered to be cycling if serum concentrations of progesterone were greater than 1 ng/ mL. For a mare to be classified as exhibiting a nocturnal rise of melatonin, serum concentrations of melatonin had to be at least two times greater at midnight than at noon. By month, a relationship did not exist (chi2; P > 0.05) among mares that were exhibiting estrous cycles and exhibiting nocturnal rises of melatonin. Likewise, examination of serum profiles of melatonin taken at 2-h intervals for 48 h revealed considerable variation among mares throughout the seasons. A nocturnal rise in serum melatonin was observed only in June (P < 0.02). In March and December, serum melatonin was greater in cycling mares than noncycling mares, but the elevation was not associated with light-dark periods (P < 0.01). Two of the mares exhibited estrous cycles throughout the seasons but melatonin secretion in these two mares were similar to that observed in the seven mares that demonstrated seasonal anestrous. From these results, it does not appear that changes in serum concentrations of melatonin are used as a cue to regulate cyclic activity in the mare throughout the seasons. (+info)
Effects of oocyte culture density on meiotic competence of canine oocytes.
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This study was conducted to determine a suitable ratio of oocytes to medium for in vitro maturation (IVM) of cumulus-oocyte complexes (COCs) collected from bitches at anoestrus and dioestrus and to examine the meiotic competence of COCs cultured singly or in different group sizes. In the first experiment, different numbers of COCs (5, 10, 15 and 20 per drop) were cultured for 72 h in 100 microl drops of maturation medium. The meiotic competence of oocytes from ovaries at anoestrus was affected by the number of COCs incubated, whereas at dioestrus, the incubation number of COCs had no effect. In the second experiment, COCs were cultured singly or in different group sizes for 72 h by suitable oocyte density according to the reproductive cycle of the donor. In the anoestrous group, 1, 5 and 10 COCs were cultured in 10, 50 and 100 microl drops of the medium (10 microl per COC), respectively. In the dioestrous group, 1, 5 and 15 COCs were cultured in 7, 35 and 105 microl drops of the medium (7 microl per COC), respectively. There were no differences in the proportions of oocytes reaching metaphase II among the different group sizes in each stage of the reproductive cycle of the donor. The results indicate that the influence of oocyte density on the meiotic competence of oocytes differs according to the stage of the reproductive cycle of the donor. Moreover, the group sizes have no effect on the meiotic competence of oocytes cultured at suitable oocyte density according to the reproductive cycle of the donor. (+info)
Follicular waves and circulating concentrations of gonadotrophins, inhibin and oestradiol during the anovulatory season in mares.
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Follicular waves and associated circulating hormone concentrations were studied during the anovulatory season in pony mares (n=8). Follicles were monitored by ultrasonography and a blood sample was taken daily from 29 January until ovulation (mean, 28 April). A mid-anovulatory period (largest follicle, 16.0+/-0.5 mm in diameter) and transitional period (largest follicle, 22.4+/-0.5 mm) were distinctive in each mare. The two periods were delineated by an increase in the diameter of the largest follicle to >/=21.0 mm. Follicular waves, identified by significant increases in the mean diameter of the second to sixth largest follicles, were detected during both the mid-anovulatory and transitional periods. The mean number of follicles >/=15.0 mm in diameter and the diameter of the second to sixth largest follicles increased in association with statistically identified FSH surges. The pattern of the FSH concentration changes during surges did not change during the mid-anovulatory and transitional periods. During the declining portion of the FSH surge, follicle growth continued and circulating total inhibin increased, indicating suppression of FSH by inhibin from the growing follicles. Circulating oestradiol or LH did not change relative to wave emergence. Results indicated that follicular waves occurred during the second-half of the anovulatory season, even during the period of lowest follicular activity. On a temporal basis, follicular wave emergence was stimulated by surges in circulating FSH. However, the increase in follicle growth to >/=21.0 mm in diameter for the wave at the beginning of the transitional period and for the subsequent waves was not attributable to a change in the characteristics of the associated FSH surges. (+info)
Ovarian cyclicity in thyroid-suppressed ewes treated with propylthiouracil immediately before onset of seasonal anestrus.
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Two experiments were conducted to determine if propylthiouracil (PTU)-induced thyroid suppression immediately before onset of anestrus would extend the breeding season in mature ewes. In Exp. 1, twice-weekly serum concentrations of progesterone indicated that all ewes were cyclic before initiation of treatment. Beginning on d 0 (January 17), ewes received 0 (n = 4), 20 (n = 5), or 40 (n = 5) mg of PTU x kg(-1) of body weight (BW) x (-1) for 35 d. Blood samples were collected regularly throughout the trial and serum thyroxine and progesterone were quantified. Ewe BW were similar (P > 0.90) among treatments before the experiment began (mean = 78.2 +/- 4.5 kg). Likewise, serum concentrations of thyroxine averaged 86.5 +/- 8.0 ng/mL on d 0. After 11 d of PTU treatment, serum thyroxine was 90.2,75.2, and 44.2 +/- 14.0 ng/mL in ewes receiving 0, 20, and 40 mg of PTU/kg BW, respectively (linear effect, P = 0.04). On d 20, thyroxine values in the three respective groups were 73.0, 51.1, and 16.1 +/- 12.9 ng/mL (linear effect, P < 0.01). Fourteen days after PTU treatment ended, serum thyroxine did not differ (P = 0.53) among the three respective groups (71.4,73.3, and 57.5 +/- 11.8 ng/mL). Ewes receiving PTU tended to weigh less on d 42 (84.2, 78.2, and 71.8 +/- 5.1 kg for ewes treated with 0, 20, and 40 mg PTU/kg, respectively; linear effect, P = 0.10). Day of onset of anestrus was designated as the day on which serum progesterone decreased and remained below 1 ng/mL. Ewes treated with 0, 20, or 40 mg of PTU/kg BW became anestrous on d 16,40, and 81 (+/- 12) of the experiment, respectively (linear effect, P < 0.01). At the time the 35-d treatment period ended, 25, 60, and 100% of ewes receiving 0, 20, or 40 mg of PTU/kg exhibited normal estrous cycles. In Exp. 2, ewes received 0, 20, or 40 mg of PTU/kg BW for 14 d. The dose was then decreased to 0, 10, and 20 mg of PTU/kg BW for the remaining 21 d. Serum thyroxine decreased to concentrations below 20 ng/mL by d 9 after initiation of PTU treatment. Ewe weights did not differ throughout the trial and no BW loss was observed. The average day that each group entered anestrus was similar to those in Exp 1. Large doses of PTU dramatically lower serum thyroxine and this effect appears to inhibit onset of anestrus in ewes. (+info)
Evidence that estrogen receptor alpha, but not beta, mediates seasonal changes in the response of the ovine retrochiasmatic area to estradiol.
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In ewes, anestrus results from a reduction in LH pulsatility due to an increased sensitivity of the hypothalamic estradiol negative feedback system. Considerable evidence has implicated the A15 group of dopaminergic neurons in the retrochiasmatic area in this seasonally dependent estradiol effect. Moreover, estradiol administered to the retrochiasmatic area in ovariectomized anestrous ewes inhibits LH secretion. However, A15 neurons do not appear to contain the classical estrogen receptors (ERalpha). Therefore, we tested the hypothesis that beta-estrogen receptors mediate the action of estradiol in the retrochiasmatic area by comparing the effects of estradiol and genistein, a selective ERbeta agonist. We also examined whether there are seasonal changes in response of the retrochiasmatic area to these agonists and if these effects are mediated by dopamine. To test these hypotheses, ovariectomized ewes were implanted with bilateral guide cannulae targeting the retrochiasmatic area. Crystalline agonists were administered via microimplants inserted down the cannulae. Blood samples taken before and 4 days after microimplant insertion were analyzed for LH concentrations, pulse frequency, and amplitude. Genistein treatment produced no significant change in LH levels in either season. Estradiol treatment decreased both mean LH concentrations and pulse frequency in anestrous but not breeding-season ewes. Administration of the dopamine antagonist sulpiride to ovariectomized ewes with estradiol microimplants in the retrochiasmatic area returned LH pulse frequency to levels indistinguishable from controls. From these data, we hypothesize that estradiol acts on local ERalpha-containing neurons in this area to stimulate a dopaminergic pathway that inhibits LH secretion during anestrus. (+info)
Effect of oestradiol treatment on mast cell populations and microflora in the vaginal cul-de-sac of seasonally anoestrous brushtail possums (Trichosurus vulpecula).
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Mast cell populations in the vaginal cul-de-sac of female brushtail possums do not appear to be related to microbial invasion but changes in their density occur at oestrus, indicating a hormonal influence. The present study examined the effect of treatment with oestradiol on microflora and on mast cell numbers and their spatial location in cul-de-sac tissue of seasonally anoestrous brushtail possums. Tissue was collected from seasonally anoestrous brushtail possums (n = 6 per group) that were either untreated (anoestrous group) or were subjected to 6 days of treatment with oestradiol (oestradiol group) administered via subcutaneous implants or with the oil vehicle alone (control group). Tissue was collected aseptically for microbiological procedures and the fractionator and optical disector were used to quantify mast cell populations. Microflora populations were low (< 4.0 x 10(4) organisms g(-1)) and numbers of mast cells were similar in all groups. Mast cell density was greatest in epithelial and connective tissues from seasonally anoestrous and control animals and lowest in oestradiol-treated possums, in which there was a significant increase in cul-de-sac mass and volume. There is an inverse relationship between circulating oestrogen concentrations and mast cell density in possum cul-de-sac tissue, which is probably the result of an increase in tissue volume. (+info)
Variation in the timing of the reproductive season among breeds of sheep in relation to differences in photoperiodic synchronization of an endogenous rhythm.
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Photoperiod may regulate seasonal reproduction either by providing the primary driving force for the reproductive transitions or by synchronizing an endogenous reproductive rhythm. This study evaluated whether breed differences in timing of the reproductive seasons of Finnish Landrace (Finn) and Galway ewes are due to differences in photoperiodic drive of the reproductive transitions or to differences in photoperiodic synchronization of the endogenous rhythm of reproductive activity. The importance of decreasing photoperiod after the summer solstice in determining the onset and duration of the breeding season was tested by housing ewes from the summer solstice in either a simulated natural photoperiod or a fixed summer-solstice photoperiod (18 h light:6 h dark; summer-solstice hold). Onset of the breeding season within each breed did not differ between these photoperiodic treatments, but Galway ewes began and ended their breeding season earlier than Finn ewes. The duration of the breeding season was shorter in Galway ewes on summer-solstice hold than on simulated natural photoperiod; duration did not differ between photoperiodic treatments in Finn ewes. The requirement for increasing photoperiod after the winter solstice for initiation of anoestrus was tested by exposing ewes from the winter solstice to either a simulated natural photoperiod or a winter-solstice hold photoperiod (8.5 h light:15.5 h dark). Onset of anoestrus within each breed did not differ between these photoperiodic treatments, but the time of this transition differed between breeds. These observations suggest that genetic differences in timing of the breeding season in Galway and Finn ewes do not reflect differences in the extent to which photoperiod drives the reproductive transitions, because neither breed requires shortening days to enter the breeding season or lengthening days to end it at appropriate times. These findings are consistent with the hypothesis that photoperiod synchronizes an endogenous rhythm of reproductive activity in both breeds and that genetic differences in timing of the breeding season reflect differences in photoperiodic synchronization of this rhythm. (+info)
Use of melengestrol acetate and gonadotropins to induce fertile estrus in seasonally anestrous ewes.
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Ewes of three genotypes (Hampshire, n = 59; Rambouillet, n = 36; crossbred, n = 57) were used to determine the efficiency of melengestrol acetate (MGA) and(or) PG-600 (a combination of pregnant mare's serum gonadotropin and human chorionic gonadotropin) in inducing fertile estrus in seasonally anestrus ewes. Ewes were assigned randomly, within genotype, to treatments in a 2 x 2 factorial arrangement. Treatments were control, .125 mg of MGA given twice per day for 9 d (MGA), a single 5-mL injection of PG-600 (PG-600), and the combination of treatments MGA and PG-600 (MGA/PG-600). Feeding of MGA began on May 14, 1990, and ended on May 23. Injections of PG-600 were given immediately after the last feeding of MGA or vehicle on May 23. All ewes were exposed to fertile, brisket-painted rams on May 24 (d 0) for 40 d. Ewes were checked for estrus twice daily for 9 d. Laparoscopy was performed, to assess ovulation rate (OR), on d 6 for ewes that were not detected in estrus and on d 12 for ewes that exhibited estrus. Percentage of ewes mated was increased by MGA (P less than .001). Ovulation rate of ewes exposed to rams was increased by PG-600 (P less than .01) and this effect was enhanced by MGA (P less than .05), whereas MGA alone tended to decrease OR (P less than .10). Melengestrol acetate decreased the interval to lambing by 6.5 d (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS) (+info)